Duan Jingyue Ellie, Jiang Zongliang Carl, Alqahtani Fahad, Mandoiu Ion, Dong Hong, Zheng Xinbao, Marjani Sadie L, Chen Jingbo, Tian Xiuchun Cindy
Department of Animal Science, University of Connecticut, Storrs, CT, United States.
School of Animal Science, AgCenter, Louisiana State University, Baton Rouge, LA, United States.
Front Genet. 2019 May 28;10:512. doi: 10.3389/fgene.2019.00512. eCollection 2019.
DNA methylation undergoes drastic fluctuation during early mammalian embryogenesis. The dynamics of global DNA methylation in bovine embryos, however, have mostly been studied by immunostaining. We adopted the whole genome bisulfite sequencing (WGBS) method to characterize stage-specific genome-wide DNA methylation in bovine sperm, immature oocytes, oocytes matured and , as well as developed single embryos at the 2-, 4-, 8-, and 16-cell stages. We found that the major wave of genome-wide DNA demethylation was complete by the 8-cell stage when methylation became prominent. Sperm and oocytes were differentially methylated in numerous regions (DMRs), which were primarily intergenic, suggesting that these non-coding regions may play important roles in gamete specification. DMRs were also identified between and matured oocytes, suggesting environmental effects on epigenetic modifications. In addition, virtually no (less than 1.5%) DNA methylation was found in mitochondrial DNA. Finally, by using RNA-seq data generated from embryos at the same developmental stages, we revealed a weak inverse correlation between gene expression and promoter methylation. This comprehensive analysis provides insight into the critical features of the bovine embryo methylome, and serves as an important reference for embryos produced , such as by fertilization and cloning. Lastly, these data can also provide a model for the epigenetic dynamics in human early embryos.
在哺乳动物早期胚胎发生过程中,DNA甲基化经历剧烈波动。然而,牛胚胎中全基因组DNA甲基化的动态变化大多是通过免疫染色进行研究的。我们采用全基因组亚硫酸氢盐测序(WGBS)方法来表征牛精子、未成熟卵母细胞、成熟卵母细胞以及发育至2细胞、4细胞、8细胞和16细胞阶段的单个胚胎的阶段特异性全基因组DNA甲基化情况。我们发现,全基因组DNA去甲基化的主要浪潮在8细胞阶段完成,此时甲基化变得显著。精子和卵母细胞在许多区域(差异甲基化区域,DMRs)存在差异甲基化,这些区域主要位于基因间,这表明这些非编码区域可能在配子特化中发挥重要作用。在未成熟和成熟卵母细胞之间也鉴定出了DMRs,表明环境对表观遗传修饰有影响。此外,在线粒体DNA中几乎未发现(少于1.5%)DNA甲基化。最后,通过使用来自相同发育阶段胚胎的RNA-seq数据,我们揭示了基因表达与启动子甲基化之间存在弱的负相关关系。这项全面分析深入了解了牛胚胎甲基化组的关键特征,并为通过体外受精和克隆等方式产生的胚胎提供了重要参考。最后,这些数据还可为人类早期胚胎的表观遗传动态变化提供模型。